Transcriptome profiling of the cysticercus stage of the laboratory model Taenia crassiceps, strain ORF

García-Montoya, Gisela María; Mesa-Arango, Jairo A.; Isaza, Juan Pablo; Agudelo‐López, Sonia P.; Cabarcas, Felipe; Barrera, Luis F.; Alzate, Juan F.

doi:10.1016/j.actatropica.2015.11.001

Cited by 13 publications

(9 citation statements)

References 48 publications

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“…The top 50 most highly represented transcripts demonstrate the expression of common conserved genes similar to other closely related cestodes at several life stages—such as dynein light chain, tegumental protein, deoxyhypusine hydroxylase, 8 kDa glycoprotein, and expressed conserved protein—thus demonstrating the validity of the transcript assembly [ 62 , 63 , 64 , 65 , 66 , 67 , 68 ]. Moreover, the A. perfoliata transcriptome aligned well to 3 closely related tapeworm species, namely H. diminuta , E. granulosus , and H. microstoma , again confirming the validity of the dataset.…”

Section: Discussionmentioning

confidence: 99%

Evidence of Immune Modulators in the Secretome of the Equine Tapeworm Anoplocephala perfoliata

et al. 2021

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Anoplocephala perfoliata is a neglected gastro-intestinal tapeworm, commonly infecting horses worldwide. Molecular investigation of A. perfoliata is hampered by a lack of tools to better understand the host–parasite interface. This interface is likely influenced by parasite derived immune modulators released in the secretome as free proteins or components of extracellular vesicles (EVs). Therefore, adult RNA was sequenced and de novo assembled to generate the first A. perfoliata transcriptome. In addition, excretory secretory products (ESP) from adult A. perfoliata were collected and EVs isolated using size exclusion chromatography, prior to proteomic analysis of the EVs, the EV surface and EV depleted ESP. Transcriptome analysis revealed 454 sequences homologous to known helminth immune modulators including two novel Sigma class GSTs, five α-HSP90s, and three α-enolases with isoforms of all three observed within the proteomic analysis of the secretome. Furthermore, secretome proteomics identified common helminth proteins across each sample with known EV markers, such as annexins and tetraspanins, observed in EV fractions. Importantly, 49 of the 454 putative immune modulators were identified across the secretome proteomics contained within and on the surface of EVs in addition to those identified in free ESP. This work provides the molecular tools for A. perfoliata to reveal key players in the host–parasite interaction within the horse host.

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Section: Discussionmentioning

confidence: 99%

Evidence of Immune Modulators in the Secretome of the Equine Tapeworm Anoplocephala perfoliata

et al. 2021

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“…Thus far, the T. crassiceps transcriptome database is the only published reference discussing GSTs in this parasite (García-Montoya et al , 2016) and was useful in supporting the characterization of TcGST1 and TcGST2. The information obtained in this study increases the utility of T. crassiceps as a model for the investigation of multifunctional GSTs.…”

Section: Discussionmentioning

confidence: 99%

“…Although T. crassiceps is an excellent experimental model to use to answer research questions concerning T. solium , the ROS detoxification mechanisms of this human parasite are poorly understood. Thus far, only one report has investigated the presence of GSTs in T. crassiceps, and enzyme functionality has not yet been evaluated (García-Montoya et al ., 2016). Given the importance of GST functionality in cestodes, and the presence of more than one enzyme isoform in all organisms, we sought to purify and characterize native GST isoenzymes in T. crassiceps , using both biochemical and kinetic procedures.…”

Section: Introductionmentioning

confidence: 99%

Two glutathione transferase isoforms isolated from juvenile cysts ofTaenia crassiceps: identification, purification and characterization

2017

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We identified and characterized the first two glutathione transferases (GSTs) isolated from juvenile cysts of Taenia crassiceps (EC 2.5.1.18). The two glutathione transferases (TcGST1 and TcGST2) were purified in a single-step protocol using glutathione (GSH)-sepharose chromatography in combination with a GSH gradient. The specific activities of TcGST1 and TcGST2 were 26 U mg-1 and 19 U mg-1, respectively, both at 25°C and pH 6.5 with 1-chloro-2,4-dinitrobenzene (CDNB) and GSH as substrates. The Km(CDNB) and Kcat(CDNB) values for TcGST1 and TcGST2 (0.86 μm and 62 s-1; 1.03 μm and 1.97 s-1, respectively) and Km(GSH) and Kcat(GSH) values for TcGST1 and TcGST2 (0.55 μm and 11.61 s-1; 0.3 μm and 32.3 s-1, respectively) were similar to those reported for mammalian and helminth GSTs. Mass spectrometry analysis showed that eight peptides from each of the two parasite transferases were a match for gi|29825896 glutathione transferase (Taenia solium), confirming that both enzymes are GSTs. The relative molecular masses were 54,000 ± 0.9 for the native enzymes and 27,500 ± 0.5 for the enzyme subunits. Thus, TcGST1 and TcGST2 are dimeric proteins. Optimal TcGST1 and TcGST2 activities were observed at pH 8.5 in the range of 20-55°C and pH 7.5 at 35-40°C, respectively. TcGST1 and TcGST2 were inhibited by cibacron blue (CB), bromosulphophthalein (BST), rose bengal (RB), indomethacin and haematin (Hm) with 50% inhibitory concentrations (IC50) in the μm range. TcGST1 was inhibited in a non-competitive manner by all tested inhibitors with the exception of indomethacin, which was uncompetitive. The discovery of these new GSTs facilitates the potential use of T. crassiceps as a model to investigate multifunctional GSTs.

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“…With the publication of the T. solium genome 12 and with the first partial cyst transcriptomes obtained by RNA-seq 13,14 ; a valuable amount of data is currently available to study stage-specific events involved in parasite’s development 15 . Moreover, whole transcriptome analysis in other tapeworms (including families Taeniidae –genuses Taenia and Echinococcus– 16–20 , Hymenolepididae 21 and Mesocestoididae 22 ) lead to the identification of conserved pathways among cestodes that compliment previous datasets.…”

Section: Introductionmentioning

confidence: 54%

Validation of Reference Genes for RT-qPCR Relative Expression Analysis in Pre-Adult Stages ofTaenia solium

Gutiérrez-Loli

Torre

Aguila

et al. 2022

Preprint

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SUMMARYThe larvae-to-adult development on the life cycle of zoonotic parasitic tapewormTaenia soliuminvolves striking -but clinically unappreciated-events with pivotal importance in cestode biology. Unlike the ones related to the intermediate host, the early-adult stages can be addressedin vitrooffering a useful model to study evagination, strobilation and worm development. In the absence of a stage-specific transcriptome, postgenomic data exploration followed by single-gene relative expression analysis by RT-qPCR (reverse transcription-quantitative PCR) are useful strategies to gather information on the regulation of genes of interest during parasite development. However, this approach requires the validation of an endogenous reference gene (RG) to achieve accurate comparisons.Therefore, we analyzed the expression stability of 17 candidate RGs on the context of the early-adult stages ofT. soliumclassified as non-evaginated and evaginated larvae (cysts). The comprehensive tool RefFinder ranked malate dehydrogenase as the most stable gene within these conditions, and its suitability for relative quantification was validated by normalizing the expression of the transporter TGTP1 gene, known for being upregulated upon evagination. This study is the first attempt in finding reliable normalization standards for transcript exploration in genus Taenia.

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Transcriptome profiling of the cysticercus stage of the laboratory model Taenia crassiceps, strain ORF

Cited by 13 publications

References 48 publications

Evidence of Immune Modulators in the Secretome of the Equine Tapeworm Anoplocephala perfoliata

Evidence of Immune Modulators in the Secretome of the Equine Tapeworm Anoplocephala perfoliata

Two glutathione transferase isoforms isolated from juvenile cysts ofTaenia crassiceps: identification, purification and characterization

Validation of Reference Genes for RT-qPCR Relative Expression Analysis in Pre-Adult Stages ofTaenia solium

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